鑑定逆境相關蛋白基因在番茄免疫途徑抗番茄黃化捲葉泰國病毒之研究

Identification of stress associated protein genes in tomato immunity against tomato yellow leaf curl Thailand virus

蔡昀珊(Yun-Shan Tsai)

指導教授 ： 葉信宏

共同指導教授 ： 沈偉強(Wei-Chiang Shen)

國立臺灣大學/生物資源暨農學院/植物病理與微生物學研究所/碩士(2022年)

https://doi.org/10.6342/NTU202200136

摘要

番茄黃化捲葉泰國病毒 (tomato yellow leaf curl Thailand virus, TYLCTHV) 對於臺灣番茄 (Solanum lycopersicum) 產業造成嚴重損害。前人研究指出雙子葉植物和單子葉植物中的逆境相關蛋白 (stress associated proteins, SAPs) 在水楊酸 (salicylic acid, SA) 所調控的抗病毒免疫途徑中扮演重要角色，為了探討番茄SAP是否具有相似的抗病毒免疫能力，本實驗目為在番茄中找出參與對抗TYLCTHV之SAPs。在模式番茄 (S. lycopersicum cv. Micro-Tom) 中進行親緣演化樹分析，確認出七個SAP基因，即SlSAP4-LIKE、SlSAP5、SlSAP5-LIKE、SlSAP8、SlSAP8-LIKE、SlSAP11和SlSAP12；其中，水楊酸可以誘導SlSAP4-LIKE、SlSAP5、SlSAP8-LIKE和SlSAP12等基因的表現量。再者，TYLCTHV的感染則可誘導SlSAP5、SlSAP8、SlSAP8-LIKE和SlSAP12的基因表現量。此外，在番茄植株中短暫過表現番茄SAP試驗發現水楊酸調控免疫相關基因受到SlSAP4-LIKE、SlSAP5、SlSAP8-LIKE和SlSAP12所調控；茉莉酸相關基因則受SlSAP8之調控；乙烯相關基因受到SlSAP8和SlSAP11的調控。另一方面，在TYLCYHV感染番茄系統中短暫過表現番茄SAP指出，SlSAP4-LIKE、SlSAP5、SlSAP8、SlSAP8-LIKE和SlSAP12可參與抗病毒免疫反應，其中，SlSAP5、SlSAP8和SlSAP8-LIKE對於減少TYLCTHV的積累有較佳的影響。此外，在in silico 啟動子分析顯示，在SlSAP5啟動子區域具有一個水楊酸反應相關的TGACG序列模體 (motif)，且SlSAP8和SlSAP8-LIKE啟動子區域則各發現一個與水楊酸反應相關的W-box序列模體。研究結果為利用SAP基因開發抗TYLCTHV策略奠定重要基礎。

關鍵字

逆境相關蛋白 ； 水楊酸 ； 番茄黃化捲葉泰國病毒 ； 番茄 ； 植物免疫

並列摘要

Tomato yellow leaf curl Thailand virus (TYLCTHV) causes severe damage to tomato (Solanum lycopersicum) production in Taiwan. Previous studies indicate that the conserved plant stress associated proteins (SAPs) serve as hubs to mediate salicylic acid (SA)-mediated antiviral immunity in both dicotyledons and monocotyledons. To analyze whether tomato SAPs have similar antiviral immunity, we try to identify tomato SAPs that confer resistance to TYLCTHV. Phylogenetic analysis identified seven SAPs, SlSAP4-LIKE, SlSAP5, SlSAP5-LIKE, SlSAP8, SlSAP8-LIKE, SlSAP11 and SlSAP12, from tomato cultivar Micro-Tom. SA treatment induced expression of SlSAP4-LIKE, SlSAP5, SlSAP8-LIKE and SlSAP12. TYLCTHV infection induced the expression of SlSAP5, SlSAP8, SlSAP8-LIKE and SlSAP12. Moreover, transient overexpression of SAPs in tomato revealed that SA-mediated immune marker genes are regulated by SlSAP4-LIKE, SlSAP5, SlSAP8-LIKE and SlSAP12; JA-related immune responsive genes are modulated by SlSAP8; ET-related immune responsive genes are regulated by SlSAP8 and SlSAP11. In addition, transient overexpression of SAPs in TYLCYHV-infected tomato indicated that SlSAP4-LIKE, SlSAP5, SlSAP8, SlSAP8-LIKE and SlSAP12 are involved in antiviral immunity. Among them, SlSAP5, SlSAP8 and SlSAP8-LIKE had more profound effects on reducing the accumulation of TYLCTHV. Furthermore, in silico promoter analysis revealed a SA responsive TGACG-motif in SlSAP5 promoter region, and a SA responsive W-box motif in SlSAP8 and SlSAP8-LIKE promoter regions. This research laid a foundation for using SAPs to develop effective strategies against TYLCTHV.

並列關鍵字

stress associated protein (SAP) ； salicylic acid ； tomato yellow leaf curl Thailand virus (TYLCTHV) ； tomato (Solanum lycopersicum) ； plant immunity

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